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Published on: December 20, 2014
Vimentin in cultured chromaffin cells: an immunofluorescent, biochemical and functional study
1Department of Physiology and Pharmacology, Centro de Ciencias Básicas,Universidad Autónoma de Aguascalientes, Aguascalientes, México. jlquinta@correo.uaa.mx
Insights
Vimentin, an intermediate filament protein, is induced in cultured adrenomedullary chromaffin cells and may regulate secretion via phosphorylation. This finding suggests a novel role for vimentin in cellular processes.
Area of Science:
- Cell Biology
- Neuroscience
- Biochemistry
Background:
- Adrenomedullary chromaffin cells are crucial for stress response, releasing catecholamines.
- The intermediate filament protein vimentin is typically absent in the adrenal medulla.
- Understanding protein dynamics in chromaffin cells is key to elucidating secretory mechanisms.
Purpose of the Study:
- To investigate the presence and function of vimentin in adrenomedullary chromaffin cells.
- To explore the regulation of vimentin by phosphorylation and its impact on cellular structure.
- To determine vimentin's potential role in catecholamine secretion.
Main Methods:
- Immunofluorescent analysis with double cell labeling using antibodies against vimentin and dopamine-beta-hydroxylase.
- Assessment of vimentin phosphorylation in response to acetylcholine and calyculin-A.
- Investigation of vimentin's role in secretion using digitonin-permeabilized cells and vimentin-specific antibodies.
Main Results:
- Vimentin expression was induced in cultured chromaffin cells following collagenase digestion.
- Vimentin phosphorylation occurred in a calcium-dependent manner, influenced by acetylcholine.
- Inhibition of vimentin phosphorylation by calyculin-A altered its distribution within the cell.
- Vimentin antibody partially inhibited calcium-induced catecholamine release, suggesting a role in secretion.
Conclusions:
- Collagenase digestion induces vimentin expression in cultured chromaffin cells.
- Vimentin phosphorylation, regulated by calcium and acetylcholine, may modulate vimentin's function.
- Vimentin appears to be involved in the regulation of the secretory process in chromaffin cells through a phosphorylation-dependent mechanism.
Abstract:
In tile present study we seek the presence and possible function of the intermediate filament protein vimentin in adrenomedullary chromaffin cells. Vimentin which is not present in the adrenal medulla was clearly showed up after collagenase digestion of the gland in the cultured chromaffin cells by using an immunofluorescent analysis with double cell labeling with monoclonal antibodies against vimentin and dopamine-beta-hydroxylase. Vimentin was also shown to be phosphorylated in a calcium-dependent manner by acetylcholine. The specific protein phosphatase inhibitor calyculin-A, that has been previously shown to increase vimentin phosphorylation, caused a change in the distribution of vimentin which moved from the Triton X-100 insoluble cytoskeletal preparation to the detergent soluble fraction probably as a result of modifications in filament integrity. The possible role of vimentin in secretion was in addition investigated using digitonin-permeabilized cells, in which the specific antibody for vimentin partially inhibited calcium-induced catecholamine release. These results demonstrate the induction of vimentin expression after collagenase digestion in cultured chromaffin cells and suggest that in these conditions this protein is possibly implicated in the regulation of the secretory process through a phosphorylation-dependent mechanism.
